Difference between revisions of "Part:BBa K3505013"

Revision as of 10:41, 26 October 2020

AndersonJ23115:LacO:RBS GB compatible A1-B2

Constitutive Anderson BBa_J23115with Lac Operator and RBS BBa_B0034

Fig.1:J23115 with LacO and RBS

Design Notes

The sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in pUPD2 BBa_K3505007 and has overhangs compatible for Golden Braid cloning. This has position A1-B3.

Fig.1:The overhangs of this part in the Golden Braid Grammar

Fig.2:The level 0 module :pupd2- AndersonJ23115:LacO:RBS I (illustration from SnapGene)

Verification of cloning

Fig.1:((U=Uncut , C= Cut) Restriction digestion of TetO (C1-C4 ) with : Not-HF(C1-C4) , Expected bands : 2046 + 129 bp ,Positive result: C1,C2,C4

Experimental Use and Experience

This part is used in BBa_K3505032

Sequence and Features

Assembly Compatibility:

10
COMPATIBLE WITH RFC[10]
12
INCOMPATIBLE WITH RFC[12]
Illegal NheI site found at 11
Illegal NheI site found at 34
21
COMPATIBLE WITH RFC[21]
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 14: / Line 14: @@
 The sequence was domesticated . We removed BsmBI ,BsaI , BtgZI, BpiI sites in order to be compatible with GoldenBraid and MoClo. The sequence is cloned in pUPD2 <bbpart>BBa_K3505007</bbpart> and has overhangs compatible for Golden Braid cloning.
 This has position A1-B3.
-[[File:T--Thessaly--GB-GGAG-AATG.jpeg|700px|thumb|none|<i><b>Fig.2:</b>The overhangs of this part in the Golden Braid Grammar</i>]]
+[[File:T--Thessaly--GB-GGAG-AATG.jpeg|700px|thumb|none|<i><b>Fig.1:</b>The overhangs of this part in the Golden Braid Grammar</i>]]
+[[File:T--Thessaly--andersonlacO-snap.png|700px|thumb|none|<i><b>Fig.2:</b>The level 0 module :pupd2- AndersonJ23115:LacO:RBS I (illustration from SnapGene)</i>]]
 ===Verification of cloning===
+[[File:T--Thessaly--TET0-digestion.png|700px|thumb|none|<i><b>Fig.1:</b>((U=Uncut , C= Cut) Restriction digestion of TetO (C1-C4 ) with : Not-HF(C1-C4) , Expected bands : 2046 + 129 bp ,Positive result: C1,C2,C4</i>]]